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研发一种非致命的快速检测方法,以准确检测欧鳗体内线虫寄生虫的存在。

Towards an non-lethal rapid test to accurately detect the presence of the nematode parasite, , in European eel, .

机构信息

Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow, G20 6NB, Glasgow, UK.

Marine Institute, Foras na Mara, F28 PF65, Newport, Ireland.

出版信息

Parasitology. 2022 Apr;149(5):605-611. doi: 10.1017/S0031182021002146. Epub 2022 Jan 19.

Abstract

Anguillicoloides crassus is an invasive nematode parasite of the critically endangered European eel, Anguilla anguilla, and possibly one of the primary drivers of eel population collapse, impacting many features of eel physiology and life history. Early detection of the parasite is vital to limit the spread of A. crassus, to assess its potential impact on spawning biomass. However accurate diagnosis of infection could only be achieved via necropsy. To support eel fisheries management we developed a rapid, non-lethal, minimally invasive and in situ DNA-based method to infer the presence of the parasite in the swim bladder. Screening of 131 wild eels was undertaken between 2017 and 2019 in Ireland and UK to validate the procedure. DNA extractions and PCR were conducted using both a Qiagen Stool kit and in situ using Whatman qualitative filter paper No1 and a miniPCR DNA Discovery-System™. Primers were specifically designed to target the cytochrome oxidase mtDNA gene region and in situ extraction and amplification takes approximately 3 h for up to 16 individuals. Our in-situ diagnostic procedure demonstrated positive predictive values at 96% and negative predictive values at 87% by comparison to necropsy data. Our method could be a valuable tool in the hands of fisheries managers to enable infection control and help protect this iconic but critically endangered species.

摘要

短沟蜷是濒危欧洲鳗的一种侵袭性线虫寄生虫,可能是导致鳗种群崩溃的主要驱动因素之一,影响鳗的许多生理和生活史特征。早期发现寄生虫对于限制短沟蜷的传播、评估其对产卵生物量的潜在影响至关重要。然而,只有通过剖检才能准确诊断感染。为了支持鳗渔业管理,我们开发了一种快速、非致死、微创和原位的基于 DNA 的方法,以推断鳔中寄生虫的存在。2017 年至 2019 年,在爱尔兰和英国对 131 条野生鳗进行了筛查,以验证该程序。使用 Qiagen Stool 试剂盒和 Whatman 定性滤纸 No1 以及 miniPCR DNA Discovery-System™ 原位提取和 PCR 进行 DNA 提取和 PCR。引物是专门设计来靶向细胞色素氧化酶 mtDNA 基因区域的,原位提取和扩增最多可对 16 个人进行约 3 小时的扩增。与剖检数据相比,我们的原位诊断程序显示阳性预测值为 96%,阴性预测值为 87%。我们的方法可以成为渔业管理者手中的有力工具,用于进行感染控制,帮助保护这种标志性但极度濒危的物种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f3fa/10268113/f6b58042ae94/S0031182021002146_figAb.jpg

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